[Internal Quality Control of Residual Pesticides Analysis Using Multiple Stable Isotope Labeled Compounds].

Internal quality control (IQC) is essential to ensure the reliability of the results of chemical analysis. In this study, we propose a novel method of IQC for multiresidue analysis of pesticides. A total of seven stable isotope labeled compounds (SILC) were added to analytical samples and were used to monitor and evaluate the quality of analytical results. In contrast to conventional IQC method in which only a limited number of control materials were analyzed to ensure the reliability of the results for an entire batch, the developed method can monitor the analytical quality of all the samples in the batch. It was shown that the developed method could achieve better performance than that of conventional method. Therefore, the developed method is considered to be promising for practical applications.(Received January 27, 2022; Accepted July 4, 2022).

Combined use of a modifier gas generator, analyte protectants and multiple internal standards for effective and robust compensation of matrix effects in gas chromatographic analysis of pesticides.

In the analysis of pesticides performed with gas chromatography, the quantitative performance of measurements can be severely compromised by phenomena known as matrix effects. In seeking a solution to the problem of matrix effects, the application of a modifier gas generator (MGG) was investigated in this study, together with analyte protectants and multiple internal standards. Ethylene glycol (EG) was used as modifier gas and matrix effects in GCMS analysis were then evaluated by using the extracts of various food commodities. MGG was used in combination with other known methods of matrix effect compensation and its performance in reducing matrix effects tested. We have found that by combining MGG with conventional analyte protectants, matrix effects were substantially reduced for most of pesticides. Use of EG was especially effective for organophosphate pesticides and those with amino groups. Using this approach, the shortcomings of conventional analyte protectants were remedied. Although neither EG nor analyte protectants could sufficiently reduce the matrix effects for certain classes of pesticides, this limitation could be overcome with the use of multiple internal standards (IS) in the analysis. Finally, it was shown that the method we developed could achieve better analytical performance than the matrix-matched calibration method. Our method was robust with respect to the variation of food matrix components, so its application to real-world analyses would be practical and promising.

Purification and characterization of cellobiose dehydrogenase from white-rot basidiomycete Trametes hirsuta.

In order to save energy during the pulp making process, we tried to use white-rot basidiomycete, Trametes hirsuta, which degrades lignin efficiently. But a decrease in paper strength caused by cellulolytic activity ruled this out for practical application. Since the cellulolytic activity of the fungus must be decreased, we purified and characterized a cellobiose dehydrogenase (CDH) that was reported to damage pulp fiber. The CDH in the culture filtrate of C. hirsutus was purified by freeze-thawing and chromatographic methods. The pI of the enzyme was 4.2 and its molecular weight was 92 kDa. The optimal temperature and pH of the enzyme were 60-70 degrees C and 5.0 respectively. Since the purified CDH decreased the viscosity of pulp in the presence of Fe(III) and cellobiose, it was shown that the suppression of CDH should be an effective way to reduce cellulose damage.

Rat cytochrome P450-mediated transformation of dichlorodibenzo-p-dioxins by recombinant white-rot basidiomycete Coriolus hirsutus.

Rat cytochrome P450, CYP1A1, has been reported to play an important role in the metabolism of mono-trichlorodibenzo-p-dioxins (M-TriCDDs). To breed lignin (and M-TetraCDDs)-degrading basidiomycete Coriolus hirsutus strains producing rat CYP1A1, an expression cassette [C. hirsutus gpd promoter-C. hirsutus gpd 5' portion (224-bp of 1st exon-8th base of 4th exon)-rat cyp1a1 cDNA-Lentinula edodes priA terminator] was constructed and inserted into pUCR1 carrying the C. hirsutus arg1 gene. The resulting recombinant plasmid, MIp5-(cyp1a1 + arg1) was introduced into protoplasts of C. hirsutus monokaryotic strain OJ1078 (Arg(-), Leu(-)), obtaining three good Arg(+) transformants. These transformants [ChTF5-2(CYP1A1), ChTF5-4(CYP1A1), and ChTF5-6(CYP1A1)] were estimated to carry nine, six, and seven copies of the expression cassette on their chromosomes, respectively. Immunoblot analysis revealed that the three transformants produce similar amounts of rat CYP1A1 enzyme. ChTF5-2(CYP1A1), ChTF5-4(CYP1A1), ChTF5-6(CYP1A1) and recipient OJ1078 were cultivated in a liquid medium containing 2,7/2,8(at a ratio of 1:1)-dichlorodibenzo-p-dioxins (2,7/2,8-DCDDs) and the amount of intra- and extracellular 2,7/2,8-DCDDs remaining was measured. The results showed that all three transformants efficiently transform 2,7/2,8-DCDDs through the action of the recombinant rat CYP1A1 enzyme.